RRR-alpha-tocopherol can be substituted for by Trolox in determination of kinetic parameters of LDL oxidizability by copper.
LDL resistance to oxidation is characterized by the lag time that precedes the rapid generation of conjugated dienes in copper-induced oxidation. Lag time (y) is described by the equation y = kx + a, where k is the efficacy constant of alpha-tocopherol (alpha-TOH), x the alpha-TOH content of LDL, and a the alpha-TOH-independent variable, all of which show considerable between-subject variability. To answer the question of whether loading of LDL with Trolox can substitute for the more timeconsuming loading with RRR-alpha-TOH in large scale studies focusing on determination of k and a values, LDL from 15 healthy subjects was loaded in vitro with 0-50 mol Trolox/mol LDL or 0-250 mM RRR-alpha-TOH. In addition, five of these subjects were supplemented in vivo with 800-1200 IU/d RRR-alpha-TOH for 17 days and k and a were determined. The ratios k(Trolox):k(alpha-TOH) and alpha(Trolox):a(alpha-TOH) were calculated. The k(Trolox) was only slightly higher than k(alpha-TOH), in contrast to alpha(Trolox), which was almost twice as high as a(alpha-TOH), indicating that, in the case of Trolox, a includes the contribution of alpha-TOH to the lag time. In the case of alpha-TOH, a includes only the additional antioxidants that occur naturally in LDL. The ratios k(Trolox):k(alpha-TOH) and alpha(Trolox):a(alpha-TOH) showed little variability, both between subjects and between in vitro and in vivo experiments. A close correlation was found between k(alpha-TOH) and a(alpha0TOH), calculated using this ratio, and k(alpha-TOH) and a(alpha-TOH), obtained in our experiments. We conclude that from this ratio, k(alpha-TOH) and a(alpha-TOH) can be reliably calculated when k(Trolox) and a(Trolox) are determined experimentally, thus allowing us to obtain k and a values in a more convenient way.
Bergmann
A R
AR
Institute of Biochemistry, Karl-Franzens University of Graz, Austria.
Ramos
P
P
Esterbauer
H
H
Winklhofer-Roob
B M
BM
D000328:Adult; D000975:Antioxidants / Q000378:metabolism; D002839:Chromans / Q000378:metabolism*; D003300:Copper / Q000494:pharmacology; D005260:Female; D006801:Humans; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000032:analysis* / Q000378:metabolism*; D008297:Male; D010084:Oxidation-Reduction; D012044:Regression Analysis; D013053:Spectrophotometry; D014810:Vitamin E / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000378:metabolism*
D000975:Antioxidants; D002839:Chromans; D008077:Lipoproteins, LDL; C099252:oxidized low density lipoprotein; D014810:Vitamin E; D003300:Copper; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid